Mir-492 Inhibits the Angiogenesis in Zebrafish (danio Rerio) Model

Purpose: Recently, it has been reported that the over expression of miR-492 exerts a potent anti-angiogenic activity in endothelial cells. In this work we investigated whether miR-492 is able to inhibit the in vivo angiogenesis. Methods: To answer the question we exploited the tumor angiogenesis zebrafish (Danio rerio) model. Briefly, either miR-492 or miR-NC (Non Coding miRNA) were transfected in cells of prostate cancer line DU-145. After 48 hours transfected cells were mixed with matrigel and samples of 1000 cells were injected (*) into the perivitelline space of more than 20 transgenic Tg(Flk1:EGFP) embryos of zebrafish in which endothelial cells express the green fluorescent protein at 48 hours post-fertilization. Within 24 hours from injection, the neovascular response originating from the developing subintestinal vessels was observed by fluorescence microscopy (arrows). Results: While xenografts of cells transfected with miR-NC allowed the vessel formation (Fig. 1a, a') xenografts of cells transfected with miR-492 cells inhibited the vessel formation (Fig. 1b, b'). Figure 1. The zebrafish angiogenesis assay Figure 1. The zebrafish angiogenesis assay Conclusions: We showed that miR-492, which impairs the angiogenic properties of endothelial cells, inhibits the development of subintestinal vessels of Danio rerio. In perspective this discloses the possibility that the zebrafish/tumor xenograft angiogenesis assay will be extensively used for testing the antiangiogenesis potential of RNA-based drugs and for investigating the tumor angiogenesis.